Method for determining by absorption of radiations the concentration of substances in absorbing and turbid matrices
First Claim
1. A method for determining and measuring in a sample matrix the concentration of an analyte which absorbs substantially monochromatic electromagnetic radiations of wavelength λ
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1, this analyte being dissolved or dispersed in the sample matrix, this method comprising the steps of;
a) selecting another electromagnetic radiation of wavelength λ
2 at which the analyte does not substantially absorb, the absorption coefficient of the matrix background being substantially the same at λ
2 and at λ
1 ;
b) by means of a source, generating a probing beam with alternant intensity controllable half-periods at λ
1 and λ
2, and splitting this beam with beam-splitter means into a reference split beam and a probe beam to be applied to the sample matrix, interaction of this probe beam and the sample matrix resulting into a response beam representative of the absorption of the probe beam by the sample matrix;
c) detecting said response beam by a first detector sensitive to both λ
1 and λ
2 thus providing a detected AC test signal and detecting the split beam with a reference detector thus providing a comparison signal;
d) optically controlling the intensity ratio of λ
1 and λ
2 half-periods using the signal from the second detector to calibrate the probing beam so that the AC signal of the first detector is essentially zero in the absence of the analyte in the sample matrix;
e) shining the calibrated probing beam to a sample matrix containing a concentration of the analyte, whereby a nonzero signal arises at the first detector, the value of this detected signal being representative of said desired determination and measurement.
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Abstract
A method and apparatus for non-invasively testing analytical substances in turbid matrices i.e. body fluids and tissues. In this method a sample is irradiated with a beam of electromagnetic energy at two alternating wavelenths, λ1 and λ2, at which the absorption by the background is the same but at one of which the radiation is absorbed by the analyte and at the other it is not. The apparatus comprises means which enable to control the input energy at λ1 and λ2 so that at the output from the sample the electric signals issuing after detection cancel in the absence of the analyte in the sample. When analyte is present cancellation no longer occurs and a signal proportional to the analyte concentration in the sample is produced. The apparatus is also designed for shifting the response back to zero when a calibrating known concentration of analyte is used as standard, thus providing a controllable zeroing base line.
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Citations
11 Claims
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1. A method for determining and measuring in a sample matrix the concentration of an analyte which absorbs substantially monochromatic electromagnetic radiations of wavelength λ
-
1, this analyte being dissolved or dispersed in the sample matrix, this method comprising the steps of;
a) selecting another electromagnetic radiation of wavelength λ
2 at which the analyte does not substantially absorb, the absorption coefficient of the matrix background being substantially the same at λ
2 and at λ
1 ;b) by means of a source, generating a probing beam with alternant intensity controllable half-periods at λ
1 and λ
2, and splitting this beam with beam-splitter means into a reference split beam and a probe beam to be applied to the sample matrix, interaction of this probe beam and the sample matrix resulting into a response beam representative of the absorption of the probe beam by the sample matrix;c) detecting said response beam by a first detector sensitive to both λ
1 and λ
2 thus providing a detected AC test signal and detecting the split beam with a reference detector thus providing a comparison signal;d) optically controlling the intensity ratio of λ
1 and λ
2 half-periods using the signal from the second detector to calibrate the probing beam so that the AC signal of the first detector is essentially zero in the absence of the analyte in the sample matrix;e) shining the calibrated probing beam to a sample matrix containing a concentration of the analyte, whereby a nonzero signal arises at the first detector, the value of this detected signal being representative of said desired determination and measurement. - View Dependent Claims (2, 3, 10)
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1, this analyte being dissolved or dispersed in the sample matrix, this method comprising the steps of;
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4. Apparatus to determine an unknown concentration of an analyte that absorbs electromagnetic radiation and is dissolved or dispersed in a sample matrix, comprising:
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a) generating means for generating an uninterrupted probe beam of the radiation which contains, alternating in time at a suitable alternating frequency, half-periods of two different and substantially monochromatic wavelengths λ
1 and λ
2 such that, at the two wavelengths, the extinction caused by the combined effects of absorption and scattering in the matrix is equal, but the absorption produced by the analyte is different;b) splitting means for splitting this beam into a reference beam to provide a reference signal to be detected by a reference detector and a test beam to provide an AC test signal to be detected by a test detector; c) optical means for transmitting the test beam into a sample of the sample matrix; d) collecting means for collecting a response of the test beam after it has traversed the sample, said collecting means comprising the test detector which is sensitive to both said wavelengths and providing in response said AC test signal; e) amplification means to amplify and rectify this AC test signal from the test detector at the wavelength-alternating frequency, to produce a DC test signal; f) compensating means for optically controlling the intensity ratio of the λ
1 and λ
2 half-periods so that the DC signal from the amplification means will be essentially zero when there is no analyte in the sample;g) control means for maintaining constant the intensity ratio of the two different wavelength half-periods of the probe beam when analyte is present in the sample using the reference signal from the reference detector; and h) display means for displaying the DC signal from the amplification means. - View Dependent Claims (5, 6, 7, 8, 9)
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11. Apparatus to determine an unknown concentration of an analyte that absorbs electromagnetic radiation and is dissolved or dispersed in a sample matrix comprising:
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a) means for generating an uninterrupted probe beam of the said radiation which contains, alternating in time, at a predetermined alternating frequency, half-periods of two different and substantially monochromatic wavelengths λ
2 and λ
2 and at the two wavelengths, the extinction caused by the combined effects of absorption and scattering of the radiation in the matrix is equal, but the absorption produced by the analyte is different;b) splitting means for splitting this probe beam into a sample beam and a reference beam and means for transmitting the sample beam into a sample of the sample matrix; c) first detector means for detecting said sample beam after it has traversed the matrix, and for generating in response an AC sample signal and means to amplify and rectify this AC sample signal to produce a DC sample signal at the wavelength-alternating frequency; d) means for optically controlling the intensity ratio of the λ
1 and λ
2 half-periods so that the signal from the test detector will be essentially zero when there is no analyte in the sample;e) second detector means for detecting said reference beam and for generating in response an AC reference signal and means to amplify and rectify this AC reference signal to generate a DC reference signal; and f) intensity control means for maintaining constant the intensity ratio of the two different wavelength half-periods of the probe beam when analyte is present in the sample using the signal from the DC reference signal, and display means for displaying the DC sample signal.
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Specification